PROJECT SUMMARY/ABSTRACT Project I A significant discovery made during the current cycle of this PPG was that effectiveness of anesthetic cardioprotection becomes attenuated in diabetic animals or in hyperglycemic conditions. We have now developed a clinically relevant model of anesthetic cardioprotection using human cardiomyocytes derived from induced pluripotent stem cells, obtained from both non-diabetic and type 2 diabetic patients. This in vitro model of human disease will enable developmental and comparative studies of normal and diabetic cardiomyocytes to address cellular and environmental mechanisms responsible for attenuation of cardioprotection efficacy in diabetics. The working hypothesis is that diabetes-related conditions increase vulnerability to stress through acute and progressive actions on mitochondria and that our strategies can reverse this diabetic phenotype. On the basis of our progress and exciting preliminary data: 1. We will determine anesthetic-induced alterations of mitochondrial bioenergetics and Ca2+ homeostasis in human ventricular cardiomyocytes (Aim 1); 2. Investigate the role of mitochondrial fission and reactive oxygen species during glucolipotoxicity (Aim 2) and; 3. Restore anesthetic cardioprotection during glucolipotoxicity in vitro and in the animal model of diabetes using pharmacological strategies (Aim 3). In summary, the cellular and molecular mechanisms that abolish volatile anesthetic cardioprotection in the diabetic heart are unknown and there are no known treatments to reverse this effect. We will focus our efforts on examination of the signaling and mitochondrial mechanisms in non-diabetic and type 2 diabetic patient-derived cardiomyocytes. Our studies will provide novel mechanistic information on the role of mitochondria and important signaling pathways that modulate cardioprotection in diabetes, and expand on the reversal of diabetic phenotype by attenuation of mitochondrial fission and reversal of nitric oxide synthase uncoupling that will be tested in a diabetic animal model.